The effective separation and analysis of pharmaceutical compounds and their metabolites is essential to researchers that develop new, more effective, safer, and economical medicines. Novel types of biologically derived drug compounds (biologics) are becoming more important and prevalent as they match and sometimes exceed the importance of traditional small synthetic molecules. However, long-established high performance liquid chromatographic (HPLC) techniques often do not provide a means to effectively separate these biologics, which are typically highly polar molecules. Recently, hydrophilic interaction liquid chromatography (HILIC) has emerged as the analytical technique of choice to analyze highly polar drugs. HILIC is perhaps the fastest growing HPLC technique, but the commercially available stationary phases may not always be able to perform all the desired/needed separations. The aim of this Phase I Small Business Innovation Research (SBIR) project is to show feasibility in using/developing a new HILIC HPLC stationary phase based on native cyclofructan. It is proposed that the cyclofructan HILIC phase may be ideal in meeting the increasing need to separate highly polar compounds of biological relevance. Initial results indicate cyclofructan in its native form may perform critical separation that existing HILIC phases cannot. It is envisioned that the unique structural features of cyclofructan (crown ether core with pendent fructofuranose units) will afford a HILIC separating agent that can perform unique separations compared to commercial HILIC columns. Further, judicious mobile phase selection may allow for the cyclofructan HILIC column to be used in different ways to separate neutral polars through cyclofructan's hydrophilic sugar moieties, cations though ion interactions with cyclofructan's crown ether, and anions by using high concentrations of cations in the mobile phase to create a cation adsorbed stationary phase that can undergo unique anion interactions. Phase II of this research will then be used to develop this technology by scaling-up column production capabilities, evaluating column and batch reproducibility, and performing beta testing with prototype columns. Ultimately, this innovative technology will be brought to market and will offer researchers the ability to achieve improved retention and selectivity of highly polar biologically relevant compounds, which is a necessity in the development of new pharmaceuticals.